Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire, UK.
University of East Anglia, Norwich, Norfolk, UK.
BMC Med Genomics. 2024 Nov 25;17(1):276. doi: 10.1186/s12920-024-02044-w.
Wound infections are a common complication of injuries negatively impacting the patient's recovery, causing tissue damage, delaying wound healing, and possibly leading to the spread of the infection beyond the wound site. The current gold-standard diagnostic methods based on microbiological testing are not optimal for use in austere medical treatment facilities due to the need for large equipment and the turnaround time. Clinical metagenomics (CMg) has the potential to provide an alternative to current diagnostic tests enabling rapid, untargeted identification of the causative pathogen and the provision of additional clinically relevant information using equipment with a reduced logistical and operative burden.
This study presents the development and demonstration of a CMg workflow for wound swab samples. This workflow was applied to samples prospectively collected from patients with a suspected wound infection and the results were compared to routine microbiology and real-time quantitative polymerase chain reaction (qPCR).
Wound swab samples were prepared for nanopore-based DNA sequencing in approximately 4 h and achieved sensitivity and specificity values of 83.82% and 66.64% respectively, when compared to routine microbiology testing and species-specific qPCR. CMg also enabled the provision of additional information including the identification of fungal species, anaerobic bacteria, antimicrobial resistance (AMR) genes and microbial species diversity.
This study demonstrates that CMg has the potential to provide an alternative diagnostic method for wound infections suitable for use in austere medical treatment facilities. Future optimisation should focus on increased method automation and an improved understanding of the interpretation of CMg outputs, including robust reporting thresholds to confirm the presence of pathogen species and AMR gene identifications.
伤口感染是一种常见的并发症,会对患者的康复产生负面影响,导致组织损伤,延迟伤口愈合,并可能导致感染扩散到伤口以外的部位。目前基于微生物检测的黄金标准诊断方法并不适合在简陋的医疗设施中使用,因为这些方法需要大型设备且周转时间长。临床宏基因组学(CMg)有可能替代当前的诊断测试,能够快速、非靶向地识别病原体,并利用具有降低后勤和操作负担的设备提供额外的临床相关信息。
本研究提出了一种用于伤口拭子样本的 CMg 工作流程的开发和演示。该工作流程应用于前瞻性收集的疑似伤口感染患者的样本,将结果与常规微生物学和实时定量聚合酶链反应(qPCR)进行比较。
伤口拭子样本在大约 4 小时内准备好进行基于纳米孔的 DNA 测序,与常规微生物学检测和物种特异性 qPCR 相比,其灵敏度和特异性值分别为 83.82%和 66.64%。CMg 还能够提供额外的信息,包括真菌种、厌氧菌、抗菌药物耐药(AMR)基因和微生物物种多样性的鉴定。
本研究表明,CMg 有可能成为一种适合在简陋医疗设施中使用的伤口感染替代诊断方法。未来的优化应侧重于提高方法的自动化程度,并更好地理解 CMg 输出的解释,包括稳健的报告阈值,以确认病原体物种和 AMR 基因鉴定的存在。